Zebularine对肝癌HepG2细胞凋亡的影响及其机制  

Effects of Zebularine on cell apoptosis in hepatocarcinoma HepG2 cells and its mechanism

作　　者：丁佑铭[1] 於展飞 廖晓峰[2] 

机构地区：[1]武汉大学人民医院肝胆腹腔镜外科,湖北武汉430060 [2]襄阳市中心医院普外科

出　　处：《胃肠病学和肝病学杂志》2015年第5期515-518,共4页Chinese Journal of Gastroenterology and Hepatology

摘　　要：目的探讨新型DNA甲基转移酶抑制剂Zebularine对肝癌细胞系Hep G2细胞凋亡的影响及其机制。方法体外培养Hep G2细胞,用不同浓度的Zebularine（25~400μmol/L）处理24、48、72 h,MTT法测定其对Hep G2细胞增殖的影响;Zebularine（100、200、400μmol/L）处理Hep G2细胞48 h后,采用流式细胞仪检测细胞凋亡,Rho123探针染色流式细胞仪检测线粒体膜电位变化,Western blotting法检测凋亡相关蛋白Bcl-2、Bax、Caspase3、Caspase9表达,并检测RUNX3基因表达情况。结果 Zebularine能抑制Hep G2细胞的增殖,呈剂量和时间依赖性;Zebularine（浓度分别为50、100、200μmol/L）作用Hep G2细胞48 h后,细胞凋亡率分别为12.4%、23.4%、35.4%,对照组凋亡率仅为3.3%;线粒体膜电位降低;凋亡相关蛋白Bax蛋白表达量升高,而bcl-2蛋白表达量降低,Pro-Caspase-3、Pro-Caspase-9表达减弱,而Cleaved-Caspase-3、Cleaved-Caspase-9表达增强,呈浓度依赖性;RUNX3表达上调,也呈剂量依赖性。结论 Zebularine能抑制Hep G2细胞的增殖并诱导其凋亡,其作用机制可能与上调RUNX3表达,进而增加细胞Casepase-3、Casepase-9活性,下调Bcl-2基因的表达及上调Bax基因有关。Objective To explore the effect of Zebularine on cell apoptosis in hepatocarcinoma HepG2 cells and its mechanism. Methods HepG2 cells were cultured in vitro. After treated by Zebularine at different concentrations re- spectively at different time, the cell survival was determined by the MTT method. Apoptosis was analyzed by flow cytom- etry. The mitochondrial membrane potential was evaluated by flow cytometry analysis after Rho123 probe staining. The changes of expression of Bcl-2, Bax, Caspase-3, Caspase-9 and RUNX3 were detected by Western blotting. Results From the data of MTT, the cell proliferation of hepatocarcinoma HepG2 cells was inhibited by Zebularine in a dose-de- pendent and time-dependent manner. Flow cytometry assays showed that Zebularine significantly induced apoptosis in HepG2 cells. After treated by Zebularine, the apoptosis rates of HepG2 cells were 12.4% , 23.4% and 35.4% , re- spectively, which showed an obvious concentration-effect relationship. Furthermore, the mitochondrial membrane poten- tial was reduced. The data of Western blotting showed that Zebularine down-regulated Bcl-2 and up-regulated Bax; the expression of Pro-Caspase-3 and Pro-Caspase-9 were decreased, and the expression of Cleaved-Caspase-3 and Cleaved- Caspase-9 were increased; and the expression of RUNX3 was increased in a dose-dependent manner. Conclusion Ze- bularine can inhibit the proliferation of HepG2 cells and induce apoptosis, and the mechanism of Zebularine on apoptosis may be related to the up regulation of RUNX3, thus down regulation the expression of Bcl-2 expression and up regulation of Bax expression, as well as the increase of activity of Caspase-3 and Caspase-9.

关 键 词：ZEBULARINE HEPG2 凋亡 RUNX3 

分 类 号：R735.7[医药卫生—肿瘤]